Holders Spain lost for the first time in 15 matches at the European Championship finals on Tuesday when Croatia beat them 2-1 to win Group D at Euro 2016.
Ivan Perisic scored the winner with three minutes left after Croatia keeper Danijel Subasic had saved a penalty from Spain captain Sergio Ramos with 20 minutes remaining.
Croatia's Nikola Kalinic had cancelled out an early goal from Spain striker Alvaro Morata in the first half.
Spain meet Group E winners Italy in the last 16 on Monday in Paris, while Croatia will play one of the third-placed teams in Lens on Saturday.
Kalinic's goal just before the break was the first goal conceded by Spain in 948 minutes of European Championship matches since October 2014.
